[go: up one dir, main page]

CN1622822A - Medicinal compositions containing ghrelin - Google Patents

Medicinal compositions containing ghrelin Download PDF

Info

Publication number
CN1622822A
CN1622822A CNA038028301A CN03802830A CN1622822A CN 1622822 A CN1622822 A CN 1622822A CN A038028301 A CNA038028301 A CN A038028301A CN 03802830 A CN03802830 A CN 03802830A CN 1622822 A CN1622822 A CN 1622822A
Authority
CN
China
Prior art keywords
acid
solution
ghrelin
buffer
pharmaceutical composition
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CNA038028301A
Other languages
Chinese (zh)
Other versions
CN100411683C (en
Inventor
南竹义春
松本大
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kangawa, Kenji
Daiichi Sankyo Co Ltd
Original Assignee
Daiichi Suntory Pharma Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Daiichi Suntory Pharma Co Ltd filed Critical Daiichi Suntory Pharma Co Ltd
Publication of CN1622822A publication Critical patent/CN1622822A/en
Application granted granted Critical
Publication of CN100411683C publication Critical patent/CN100411683C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/27—Growth hormone [GH], i.e. somatotropin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/25—Growth hormone-releasing factor [GH-RF], i.e. somatoliberin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/04—Drugs for genital or sexual disorders; Contraceptives for inducing labour or abortion; Uterotonics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/12—Drugs for genital or sexual disorders; Contraceptives for climacteric disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/02—Drugs for disorders of the endocrine system of the hypothalamic hormones, e.g. TRH, GnRH, CRH, GRH, somatostatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/06—Drugs for disorders of the endocrine system of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/10—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
    • A61P5/44—Glucocorticosteroids; Drugs increasing or potentiating the activity of glucocorticosteroids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Endocrinology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Diabetes (AREA)
  • Immunology (AREA)
  • Neurology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Zoology (AREA)
  • Neurosurgery (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biomedical Technology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Dermatology (AREA)
  • Molecular Biology (AREA)
  • Rheumatology (AREA)
  • Psychiatry (AREA)
  • Reproductive Health (AREA)
  • Biochemistry (AREA)
  • Virology (AREA)
  • Inorganic Chemistry (AREA)
  • Pregnancy & Childbirth (AREA)
  • Urology & Nephrology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Hematology (AREA)

Abstract

本发明目的在于提供一种含有稳定态的生长素释放肽(生长素)或生长素释放肽衍生物的药物组合物，生长素释放肽是促生长激素分泌受体(GHS－R)的内源性促生长激素分泌剂(GHS)。该含生长素释放肽的药物组合物包括水溶液，其特征在于该溶解有生长素释放肽的水溶液的pH值为2－7，其中该pH值为2－7的水溶液是缓冲溶液，具体地说是甘氨酸盐酸盐缓冲液、醋酸盐缓冲液、柠檬酸盐缓冲液、乳酸盐缓冲液、磷酸盐缓冲液、柠檬酸－磷酸盐缓冲液、磷酸盐－醋酸盐－硼酸盐缓冲液或邻苯二甲酸盐缓冲液，生长素释放肽或其衍生物在该水溶液中的浓度为0.03nmol/mL－6μmol/mL。

Figure 03802830

The object of the present invention is to provide a pharmaceutical composition containing stable ghrelin (ghrelin) or ghrelin derivatives, ghrelin is the endogenous source of growth-stimulating hormone secretion receptor (GHS-R) Sexual growth hormone secretor (GHS). The ghrelin-containing pharmaceutical composition includes an aqueous solution, which is characterized in that the aqueous solution in which ghrelin is dissolved has a pH value of 2-7, wherein the aqueous solution with a pH value of 2-7 is a buffer solution, specifically Glycine hydrochloride buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer, phosphate-acetate-borate buffer solution or phthalate buffer solution, the concentration of ghrelin or its derivatives in the aqueous solution is 0.03nmol/mL-6μmol/mL.

Figure 03802830

Description

The pharmaceutical composition that contains ghrelin
Technical field
The present invention relates to a kind of ghrelin of stable state or pharmaceutical composition of ghrelin derivant of containing, ghrelin is the endogenous growth hormone succagoga (GHS) of secretagogue receptor (GHS-R), the invention still further relates to a kind of modification hydrophobic group of ghrelin or derivatives thereof that prevents the method for degraded takes place in the aqueous solution that is dissolved with the ghrelin or derivatives thereof.
Background technology
Ghrelin (auxin) is a kind of orphan receptor---the endogenous growth hormone succagoga (GHS) of secretagogue receptor (GHS-R), it be early than 1999 by rat separates and purification obtains a kind of physiologically active peptide (Kojima etc., Nature, 402:656-660,1999).After this, for example the mankind, rat, pig, chicken, Anguillar japonica, cattle, horse, sheep, the frog, salmon and Canis familiaris L. also separate and obtain the multiple ghrelin that has with the identical chemical constitution of rat ghrelin by other vertebrates beyond the rat.The chemical constitution of these ghrelins is listed in the table below in 1.
Table 1
Human mouse rat pig Niu Mianyang dog eel salmon chicken bullfrog Tilapia mossambica catfish horse GSS (positive caprylyl) FLSPEHQRVQQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPEHQRVQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPEHQKAQQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPEHQKAQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPEHQKAQQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPEHQKVQQRKESKKPAAKLKPR GSS (positive caprylyl) FLSPEHQKLQRKEAKKPSGRLKPR GSS (positive caprylyl) FLSPEHQKLQRKEPKKPSGRLKPR GSS (positive caprylyl) FLSPEHQKLQQRKESKKPPAKLQPR GSS (positive caprylyl) FLSPSQRPQGKDKKPPRV-NH2GSS (positive caprylyl) FLSPSQKPQVRQGKGKPPRV-NH 2GSS (positive caprylyl) FLSPSQKPQGKGKPPRV-NH 2GSS (positive caprylyl) FLSPTYKNIQQQKGTRKPTAR GSS (positive caprylyl) FLSPTYKNIQQQKDTRKPTAR GSS (positive caprylyl) FLSPTYKNIQQQKDTRKPTARLH GLT (positive caprylyl) FLSPADMQKIAERQSQNKLRHGNM GLT (positive caprylyl) FLSPADMQKIAERQSQNKLRHGNM GLT (positive caprylyl) FLSPADMQKIAERQSQNKLRHGNMN GSS (positive caprylyl) FLSPSQKPQNKVKSSRI-NH2GSS (positive caprylyl) FLSPTQKPQNRGDRKPPRV-NH 2GSS (positive caprylyl) FLSPTQKPQNRGDRKPPRVG GSS (positive caprylyl) FLSPEHHKVQHRKESKKPPAKLKPR
(wherein, amino acid residue is write according to IUPAC and the defined one-letter symbol of IUC)
These peptides are characterised in that owing to the ad hoc structure that hydroxyl is for example sad by fatty acid or the capric acid acidylate produces on its serine group (S) or threonine group (T) side chain, do not isolate for example this physiologically active peptide with modification property hydrophobic group of ghrelin as yet.These new peptides have the excretory effect of effective somatropin, and clear these peptides of recognizing can be regulated secretion of growth hormone.Therefore, a lot of research worker are for the physiologically active form of ghrelin and how with these polypeptide exploitation becoming medicines (for example open WO 01/07475 of world patent) quite interested.
It is considerable (Kojima etc., Nature, 402:656-660,1999) that modification hydrophobic group in the known ghrelin molecule has physiologically active for it.Yet, since do not exist resemble ghrelin this in molecule the hydroxyl place on special acid residue side chain have the peptide of modification property hydrophobic group, therefore also never the stability of these peptides was carried out research so that it is developed to medicine.
Incidentally, the chemical compound that can be developed into to medicine has various types of chemical constitutions, and just because of these chemical constitutions make this compounds may after the preparation process in or in storage process, be easy to degrade.This class degradation reaction has hydrolytic rupture, dehydration, isomerization, cancellation, oxidation, reduction or the light degradation of chemical compound, and these chemical compounds and with the additive generation chemical reaction of its compatibility.Therefore, for these chemical compounds being developed to medicine and subsequently it is carried out quality control, to the degradation reaction type that causes by the chemical constitution of these chemical compounds and that palliating degradation degree is studied and understanding seems is extremely important.
As everyone knows, stability of drug is subjected to the influence of surrounding to a great extent, for example the pH level of environment.Studied the influence of pH value of solution, and also reported the pH curve (for example SumieYoshioka, " Stability of Medicines " published 1995 by Nankohdo) of the degradation speed of a lot of medicines for the degradation speed of water-soluble liquid drug.
Physiologically active peptide or physiologically active protein matter can be present in the protease deactivation in the digestive organs and degrade, so are difficult to develop and contain these peptides or proteinic composition for oral administration.So for clinical administration, these peptides or protein are made into the form of injectable composition, yet in order to reach this purpose, this class material the stability in the aqueous solution for be made into liquid pharmaceutical formulation (though be which kind of dosage form for example the solution form or can be in the dissolved solution form of site) for all seem extremely important.
At present, contain various types of polypeptide or protein for example the pharmaceutical composition of short natruresis peptide, LH-RH (LHRH) derivant or thyroliberin derivant of insulin, growth hormone, calcitonin, atrium go on the market as medicine, it is reported that these peptide classes or proteinic chemical change have desamidization, the formation of different aspartic acid, hydrolytic rupture is for example cracked, racemization, formation two sulfur valence link or exchange reactions, β-elimination or oxidation reaction.
These chemical changes can influence the stability that this class contains peptide or protein compositions, and the degree that peptide class or protein take place to degrade depends on the pH value of solution.For example, it is reported that amount that the chemical constitution of catabolite and catabolite produce is with containing these peptide class or protein, LH-RH derivant (Strickley etc., Pharm.Res., 7 for example, 530-536,1990), human parathryoid hormone (Nobuchi etc., Pharm.Res., 14,1685-1690,1997), (antithrombotic forms material: Gietz etc., Pharm.Res., 15 to hirudin, 1456-1462,1998) and human amylin derivatives (Hekmann etc., Pharm.Res., 15, the pH value of solution 650-659,1998) changes and changes.
Ghrelin or derivatives thereof of the present invention is a kind of physiologically active peptide, and the aqueous solution that normally will contain ghrelin is made the pharmaceutical composition that is used for medicine.Although the stability of ghrelin in aqueous solution is extremely important for preparation of drug combination, the stability of ghrelin in aqueous solution was not carried out any research as yet so far.Have special modification hydrophobic group in the molecule of ghrelin or derivatives thereof, that is exactly that hydroxyl on the side chain of some amino acid residue of ghrelin or derivatives thereof is by the fatty acid acidylate.Also never find to resemble this peptide that in molecule, has special modification hydrophobic group of ghrelin, therefore also never reported the general knowledge of relevant ghrelin stability.That is to say that the mechanism of the chemical constitution of present stability for the ghrelin catabolite, catabolite and catabolite degraded is still not clear.In addition, degrade also unclear for the mechanism of degradation of ghrelin modification property hydrophobic group and the secondary of ghrelin catabolite.
Under such background, the object of the present invention is to provide a kind of pharmaceutical composition that can stably contain the ghrelin or derivatives thereof, and the present invention also aims to provide a kind of modification hydrophobic group of ghrelin or derivatives thereof that prevents that the method for degraded takes place in the aqueous solution that is dissolved with the ghrelin or derivatives thereof, this method is carried out the knowledge that institute obtains based on the chemical stability to the ghrelin or derivatives thereof that has special modification hydrophobic group in the molecule.
By the relation of the influence between the chemical constitution of the pH that contains the ghrelin aqueous solution and ghrelin catabolite is furtherd investigate; the present inventor finds; ghrelin is degraded to produce and is taken off acylated compound owing to its special modification hydrophobic group generation hydrolytic rupture in aqueous solution; simultaneously it is also because β-elimination reaction generation dehydroalanine chemical compound of degrading takes place in its modification property hydrophobic group; because the dehydroalanine chemical compound has volatility and will produce the secondary catabolite, all these degradation reactions all are subjected to the influence of pH value of aqueous solution then.
On the result of study basis of above-mentioned ghrelin mechanism of degradation, the present inventor further finds can stably be contained by the pH that uses pH regulator agent or buffer agent regulator solution the pharmaceutical composition of ghrelin, and by using different types of buffer agent no matter how its concentration or grhelin concentration can reach this stablizing effect, thereby finished the present invention.
Disclosure of the Invention
Therefore, an aspect of of the present present invention is to provide a kind of ghrelin or derivatives thereof pharmaceutical composition of (hereinafter and in claims being called " ghrelin ") that contains, and the pH that wherein is dissolved with the aqueous solution of this ghrelin is 2-7.
More particularly, the invention provides following aspect:
(1) contain the pharmaceutical composition of ghrelin, the pH that wherein is dissolved with the aqueous solution of this ghrelin is 2-7.
(2) according to the pharmaceutical composition of (1), wherein said pH is 3-6.
(3) according to the pharmaceutical composition of (1) or (2), wherein also contain pH regulator agent or buffer agent.
(4) according to the pharmaceutical composition of (3), wherein the pH regulator agent is selected from following one or more: hydrochloric acid, sulphuric acid, nitric acid, boric acid, carbonic acid, heavy carbonic, gluconic acid, sodium hydroxide, potassium hydroxide, ammonia, citric acid, monoethanolamine, lactic acid, acetic acid, succinic acid, fumaric acid, maleic acid, phosphoric acid, pyrovinic acid, malic acid, propanoic acid, trifluoroacetic acid and their salt.(5) according to the pharmaceutical composition of (3), wherein buffer agent is selected from following one or more: glycine, acetic acid, citric acid, boric acid, phthalic acid, phosphoric acid, succinic acid, lactic acid, tartaric acid, carbonic acid, hydrochloric acid, sodium hydroxide and their salt.
(6) according to a kind of pharmaceutical composition of one of (3)-(5), wherein pH regulator agent or the buffer agent concentration in solution is 0.01mM-1000mM.
(7) according to a kind of pharmaceutical composition of one of (1)-(6), wherein solution is buffer solution.
(8) according to the pharmaceutical composition of (7), wherein buffer solution is glycine hydrochloride salt buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer, phosphate-acetate-borate buffer solution or phthalate buffer.
(9) according to a kind of pharmaceutical composition of one of (1)-(8), wherein the concentration of ghrelin in solution is 0.03nmol/mL-6 μ mol/mL.
(10) according to a kind of pharmaceutical composition of one of (1)-(9), wherein ghrelin is an acetate.
(11) according to a kind of pharmaceutical composition of one of (1)-(10), wherein ghrelin is the plain release peptide of human growth.
(12) according to a kind of pharmaceutical composition of one of (1)-(11), wherein also contain anti-adsorbent.
(13) according to the pharmaceutical composition of (12), wherein the concentration of anti-adsorbent is 0.001%-5%.
(14) according to the pharmaceutical composition of (12) or (13), wherein anti-adsorbent is a surfactant.
(15) contain the pharmaceutical composition of ghrelin, wherein contain the powder that a kind of solution drying by one of (1)-(14) obtains.
(16) according to the pharmaceutical composition of (15), wherein powder is a freeze-dried powder.
(17) prevent that the hydrophobic group of ghrelin from the method for degraded taking place in containing the solution of ghrelin, it comprises that the pH of regulator solution is 2-7.
(18) according to the method for (17), wherein the described pH of regulator solution is 3-6.
(19) method of basis (17) or (18) is comprising pH regulator agent or buffer agent.
(20) method of basis (19) is comprising being selected from one or more following pH regulator agent: hydrochloric acid, sulphuric acid, nitric acid, boric acid, carbonic acid, heavy carbonic, gluconic acid, sodium hydroxide, potassium hydroxide, ammonia, citric acid, monoethanolamine, lactic acid, acetic acid, succinic acid, fumaric acid, maleic acid, phosphoric acid, pyrovinic acid, malic acid, propanoic acid, trifluoroacetic acid and their salt.
(21) method of basis (19) is comprising being selected from one or more following buffer agents: glycine, acetic acid, citric acid, boric acid, phthalic acid, phosphoric acid, succinic acid, lactic acid, tartaric acid, carbonic acid, hydrochloric acid, sodium hydroxide and their salt.
(22) according to a kind of method of one of (19)-(21), pH regulator agent or the buffer agent concentration in solution is 0.01mM-1000mM.
(23) according to a kind of method of one of (17)-(22), wherein solution is buffer solution.
(24) according to the method for (23), wherein buffer solution is glycine hydrochloride salt buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer, phosphate-acetate-borate buffer solution or phthalate buffer.
(25) according to a kind of method of one of (17)-(24), wherein the concentration of ghrelin in solution is 0.03nmol/mL-6 μ mol/mL.
(26) according to a kind of method of one of (17)-(25), wherein ghrelin is an acetate.
(27) according to any one method of (17)-(26), wherein ghrelin is the plain release peptide of human growth.
The accompanying drawing summary
Fig. 1 has shown the HPLC figure about the degraded mode of deacylation compound in the ghrelin aqueous solution of embodiment 1 and Dha chemical compound.
Fig. 2 is the diagram chart that shows the pH-degradation kinetics constant curve of ghrelin.
Fig. 3 is the diagram chart that is presented at the catabolite growing amount of the ghrelin in the aqueous solution with dissimilar pH.
Fig. 4 is the diagram chart that is presented at the pH stability of the ghrelin in the dissimilar buffer solution.
Finish optimum implementation of the present invention
To more specifically explain pharmaceutical composition of the present invention below.
Ghrelin used in the present invention is the endogenous growth hormone secretagogues, and this peptide has the effect that improves intracellular calcium concentration and induced growth hormone secretion.Suitable ghrelin is that those are obtained by the mankind, rat, pig, chicken, Anguillar japonica, cattle, horse, sheep, the frog, salmon or Canis familiaris L..In the present invention, the preferred ghrelin that is obtained by the mankind that uses more preferably uses obtain and the plain release peptide of human growth that have 28 amino acid residues by the mankind.
Modification hydrophobic group as one of ghrelin feature is not limited in caprylyl (C 8), it can be for having 2-20, the fatty acid residue of preferred 4-12 carbon atom, for example caproyl (C 6), capryl (C 10) or lauroyl (C 12).In addition, hydrophobic group can be for the side chain residue of saturated or unsaturated fatty acid, have for example fatty acid residue of hydrocinnamoyl and diamantane (obsolete) skeleton of aryl.
Therefore, ghrelin derivant of the present invention comprises the polypeptide that is set forth in the above-mentioned table 1, by inserting, increase and deleting one or more aminoacid and/or, can also modify with chemical method if necessary by modifying aminoacid sequence wherein with the described aminoacid sequence of other aminoacid replacement.In addition, ghrelin derivant of the present invention also comprises such peptide class, and wherein modification property hydrophobic group also has the physiologically active identical with ghrelin by ester bond with amino acid chain connection and it.
The ghrelin that is used for pharmaceutical composition of the present invention comprises the peptide class and the salt thereof of free form.Can transform mutually between the peptide of free form and its salt.The peptide of free form is by being converted into pharmaceutically-acceptable salts with inorganic or organic acid reaction.The example of this salt comprises inorganic acid salt for example carbonate, heavy carbonate, hydrochlorate, sulfate, nitrate or borate, and acylate for example succinate, acetate, propionate or trifluoroacetate.Also comprise alkali metal salt for example sodium salt or potassium salt in addition, alkali salt is calcium salt or magnesium salt for example, and the salt of organic amine is triethylamine salt for example, and the salt of basic amino acid alginate for example.Peptide class of the present invention can with metal complex for example the form of copper complex or Zn complex exist.
The form of above-mentioned salt has important function for the stability of ghrelin.That is to say that because the pH value of water solution of above-mentioned salt has nothing in common with each other, so these salt play a part the pH regulator agent for the water-soluble ripple of ghrelin.
The source of ghrelin and preparation method are not limited by the present invention.Can use in the present invention by the combination of chemical method, quasi-chemical method, genetic method or these methods and the ghrelin that obtains by the organism extraction.
Ghrelin as pharmaceutical raw material normally provides in the form with freeze-dried powder after purification such as reversed-phase liquid chromatography.
Aqueous solution of the present invention or solution are the solution that obtains as solvent with water, however the solvent that within pharmaceutically acceptable scope, also can use other for example ethanol, 2-propanol etc.
The concentration of ghrelin in pharmaceutical composition of the present invention is also unrestricted, preferably within pharmaceutically acceptable scope.Concentration limit is ghrelin has physiologically active under this concentration a concentration, and upper limit of concentration is that ghrelin can be dissolved in the concentration in the aqueous solution under this concentration.As for example preferred usually 0.03nmol/mL-6 μ mol/mL of the concentration of medicine, more preferably use the concentration of 0.03nmol/mL-3 μ mol/mL.
In the acceptable composition, the pH value of solution is 2-7, more preferably 3-6 on the physiology that can stably contain ghrelin of the present invention.The stable pH value that discovery contains the solution of ghrelin is 2-7.Use pH regulator agent or buffer agent to regulate the pH value of solution that this contains ghrelin.
The example of pH regulator agent comprises hydrochloric acid, sulphuric acid, nitric acid, boric acid, carbonic acid, heavy carbonic, gluconic acid, sodium hydroxide, potassium hydroxide, ammonia, citric acid, monoethanolamine, lactic acid, acetic acid, succinic acid, fumaric acid, maleic acid, phosphoric acid, pyrovinic acid, malic acid, propanoic acid, trifluoroacetic acid and their salt.
The example of buffer agent comprises glycine, acetic acid, citric acid, boric acid, phthalic acid, phosphoric acid, succinic acid, lactic acid, tartaric acid, carbonic acid, hydrochloric acid, sodium hydroxide and their salt.Wherein, preferred glycine, acetic acid or succinic acid are as buffer agent.
Consider the stability of ghrelin in aqueous solution, more satisfactory situation is to reduce the fluctuation of solution pH value.Therefore, pharmaceutical composition of the present invention is the solution with buffer capacity, just buffer solution.
Use has the buffer solution that can suppress the ghrelin degraded under its pH scope, thereby uses pH to be 2-7, more preferably the solution of 3-6.Suitable buffer solution is glycine hydrochloride salt buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer (comprising the Mcllvaine buffer), phosphate-acetate-borate buffer solution (comprising the Britton-Robinson buffer) and phthalate buffer.The example of composition comprises above-mentioned buffer agent in each buffer.
The concentration of pH regulator agent is unrestricted, and it can use the concentration of 0.01-100mM usually for being generally used for the concentration that regulator solution obtains required pH scope.
In addition, the concentration of buffer agent is also unrestricted, and it can be for keeping the concentration of buffer capacity.Generally speaking, use 0.01-100mM usually, preferred 0.1-100mM, the more preferably concentration of 1-100mM.
According to the present invention, provide the pharmaceutical composition that can stably contain the aqueous solution form of ghrelin.Consider the low irritant and the antiseptic effect of Morie osmolarity, dissolubility, solution and prevent that solution composition is absorbed, this pharmaceutical composition can also contain other additive.
Thereby can worry usually that peptide class or protein can be attracted in the preparation process of the pharmaceutically acceptable water solution that contains them and administration process reduces peptide class or proteinic concentration on the used container.In the situation of pharmaceutical composition of the present invention, confirmed that the amount that ghrelin is adsorbed on glass container or the polypropylene containers falls in the concentration range of its hyoscine.Therefore, preferably adding anti-adsorbent is adsorbed by container to prevent ghrelin.The example of anti-adsorbent comprises surfactant, saccharide, aminoacid and protein.
Surfactant of the present invention comprises surfactant cited in " handbook of pharmaceutical excipients " and the chemical compound with surface activity effect, suitable surfactant is selected from these surfactant examples, comprises quaternary ammonium salt, polyoxyethylene sorbitol fatty acid ester, Sorbitol fatty acid ester, p-Hydroxybenzoate, Polyethylene Glycol, phospholipid, bile acid, polyoxyethylene castor oil class, polyoxyethylene, polyoxyethylene polyoxypropylene, polyhydric alcohol, anion surfactant, synthetic or semi synthetic polymer.Wherein, preferably use polyoxyethylene sorbitol fatty acid ester and Sorbitol fatty acid ester.
Suitable quaternary ammonium salt comprises benzalkonium chloride, Benzethonium Chloride and hexadecane base pyridinium chloride.
Suitable polyoxyethylene sorbitol fatty acid ester comprises polyoxyethylene sorbitol monolaurate (polysorbate 20 or tween 20), polyoxyethylene sorbitol monopalmitate (polysorbate 40 or tween 40), polyoxyethylene sorbitol monostearate (polysorbate 60 or tween 60), polyoxyethylene sorbitol tristearate (polysorbate 65 or tween 65), polyoxyethylene sorbitol monoleate (polysorbate 80 or tween 80) and polyoxyethylene sorbitol trioleate (polysorbate 85 or tween 85).
Suitable Sorbitol fatty acid ester comprises the sorbityl monododecanoate (class of department 20), Sorbitol monopalmitate (span 40), Sorbitol monostearate (class of department 60), sorbitol monooleate (class of department 80), Sorbitol trioleate (class of department 85) and the Sorbitol sesquioleate.
Suitable p-Hydroxybenzoate comprises methyl parahydroxybenzoate, ethylparaben, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate and p-Hydroxybenzoic acid isobutyl ester.
Suitable Polyethylene Glycol comprises ethylene glycol furfural (ethylene glycol furfural 75), Mcrogol 400 (PEG400s), Mcrogol 600 (Macrogol 600) and Mcrogol 4000 (polyethylene glycols 4000); Suitable phospholipid comprises purified soybean lecithin and purified yolk lecithin; Suitable bile acid comprises deoxidation bile acid sodium.
Suitable polyoxyethylene castor oil class comprises polyoxyethylene castor oil, polyoxyethylene hydrogenated Oleum Ricini, polyoxyethylene hydrogenated Oleum Ricini 50 and polyoxyethylene hydrogenated Oleum Ricini 60.Other polyoxyethylene example comprises polyoxyl 10 oleyl ether, polyoxyethylene octadecyl ether, polyoxyethylene hexadecane base ether and polyoxyethylene lauryl sulfate.
Suitable polyoxyethylene polyoxypropylene comprises polyoxyethylene polyoxypropylene ethylene glycol (pluronic ) and polyoxyethylene polyoxypropylene hexadecane base ether.
Suitable polyhydric alcohol comprises glycerol (glycerol), propylene glycol and single glyceryl stearate; Suitable anion surfactant comprises alkyl ether sulfate for example hexadecane base sodium sulfate, 12 carbon alkyl sodium sulfates and oil base sodium sulfate; Sulfo-succinic acid alkyl ester salt is sulfo-succinic acid dodecyl ester sodium for example.Suitable synthetic or semi synthetic polymer comprises polyvinyl alcohol, CVP Carbopol ETD2050, polyvinylpyrrolidone and sodium polyacrylate.
The example of saccharide comprises monosaccharide for example mannitol, glucose, fructose, inositol, Sorbitol and xylitol; Disaccharide is lactose, sucrose, maltose and trehalose for example; Polysaccharide is starch, glucosan, amylopectin, alginic acid, hyaluronic acid, pectic acid, phytic acid, phytin, chitin and chitosan for example.The example of dextrin comprises alpha-cyclodextrin, beta-schardinger dextrin-, gamma-cyclodextrin, dextrin, hydroxypropyl starch and hydroxyl starch.Cellulosic example comprises methylcellulose, ethyl cellulose, hydroxyethyl-cellulose, hydroxypropyl cellulose and hydroxypropyl emthylcellulose, sodium carboxymethyl cellulose.
Suitable aminoacid comprises glycine and taurine; And polyamino acid for example polyglutamic acid, poly-aspartate, polyglycine and poly-leucine.Proteinic example comprises albumin and gelatin.
When pharmaceutical composition of the present invention is used as detectable or veterinary drug, can use the anti-adsorbent of non-human serum albumin as said composition; Yet, when said composition is used as the medicine for the treatment of human diseases, preferred end user's serum albumin.
These anti-adsorbents can be united use.The concentration of anti-adsorbent is such scope, and wherein the consumption of this anti-adsorbent is the pharmaceutically acceptable consumption while can also suppress ghrelin to be adsorbed by container, and various composition can not assembled in preparation process or long term storage process.For example, the concentration of anti-adsorbent is 0.001-5%, is preferably 0.01-1%.
Pharmaceutical composition of the present invention can also contain the additive that is useful on any purpose, and the example of additive is selected from " handbook of pharmaceutical excipients 2000 " (Japanese drug excipient committee: YakujiNippoh Sha).These additives comprise isotonic agent for example sodium chloride and mannitol; Antiseptic is sodium benzoate for example; Antioxidant is sodium sulfite, sodium pyrosulfite and ascorbic acid for example; Placebo is lidocaine hydrochloride and Mepivacaine Hydrochloride for example.
Pharmaceutical composition of the present invention can be prepared according to the used conventional method of pharmaceutical field.For example, at first freeze dried ghrelin is dissolved in and purifies waste water, also buffer agent, anti-adsorbent and other additive are dissolved in simultaneously during another part purifies waste water.Merge resulting these aqueous solutions then, if necessary can filtration sterilization, the solution that obtains is loaded on the pharmaceutical composition that has just obtained containing ghrelin of the present invention in ampoule or the bottle.
As for the dosage form of ejection preparation, can select the on-the-spot preparation of this pharmaceutical composition.This dosage form is suitable for unstable compounds in the solution of long term storage.Therefore, being used for this compositions that contains the solution of ghrelin of on-the-spot preparation is one of ejection preparation of the present invention.Be used for the solid additive that this compositions that contains the solution of ghrelin of on-the-spot preparation can contain medicinal ghrelin raw material and other necessary amounts.In addition, said composition is to obtain by this solution that contains the additive of ghrelin and other necessary amounts is carried out drying.The dry technology of solution can be freeze-drying method or spray drying process, the preferably freeze drying method.Such solid composite can the scene be dissolved in the water and uses as solution.
Pharmaceutical composition of the present invention can be used as medicine to mammal (people, monkey, Canis familiaris L., rat etc.) administration.Disease that said composition is suitable for or accessible effect comprise that those and growth hormone (GH) lack or reduce diseases associated, for example the osteoblast of cretinism, activation normal adult or osteanagenesis, raising muscle quantity and muscle strength, improvement lack the serious schizophrenia of adult's physical ability, the treatment childhood of GH, with promoting sexual gland hormone unite use with induced spawning, prevent to take the protein metabolism that prednisone causes not normal, accelerate the T cell culture in serious immune deficiency disorder, the advanced age weightlessness and prevent fatty surplus and atrophoderma.
In addition, have the effect that improves heart rate based on pharmaceutical composition of the present invention, those directly lack with growth hormone (GH) or reduce relevant suitable disease and can expect that maybe the example of the effect that reaches also comprises for example heart failure of cardiovascular disease.The effect of pharmaceutical composition of the present invention is not limited to the people, also can be used to promote animal growth, reduce fat etc., and these effects are more obvious than the effect of taking the GH acquisition.Because pharmaceutical composition of the present invention can improving appetite, therefore it can be used as the appetite improving agent of treatment anorexia or anorexia nervosa by administration in intravenous or the brain ventricle.In addition, pharmaceutical composition of the present invention can also be used for the treatment of the not normal for example non-ucler dyspepsia of gastric motility, spontaneous moderate gastroatonia, dynamic property dyspepsia and adverse current esophagitis.
In addition; pharmaceutical composition of the present invention can be accelerated in the bone marrow, duodenum and the growth of the cell in the jejunum, so its protective agent, prevention of can be used as intestinal mucosa causes the medicament of mucous membrane of small intestine damage and is used for prevention of osteoporosis disease in intravenous is supplied with the process of nutrition.
In addition, pharmaceutical composition of the present invention can be used for treating following disease or improves following body function.The example of these diseases comprises the release that stimulates old people's growth hormone, the metabolic side effect of prevention glucocorticoid, treatment and prevention of osteoporosis disease, stimulating immune system, promote the treatment of damage, promote the reparation of fracture, the treatment growth retardation, renal failure or functional defect that treatment is caused by growth retardation, treatment physiological function disappearance symptom comprises the not enough and symptom relevant with chronic disease of children growth hormone, treatment is with the growth retardation or the growth retardation relevant with obesity of treatment of obesity, treat the growth retardation relevant with Turner's syndrome with the Prader-Willi syndrome, promote the recovery and the minimizing of burn to be in hospital, developing womb is slow, skeleton development is unusual, treatment hypercorticoidism and Cushing's syndrome, induce the release of cardiac contractility growth hormone, nervous patient's growth hormone succedaneum, dyschondroplasia, Noonan ' s syndrome, schizophrenia, depression, degenerative brain disorder, treatment postpones damage and treatment social mentality disappearance, treatment pulmonary insufficiency and breathing dependence syndrome, the protein metabolism reaction decays after postponing major operation, protein loss and cachexia that minimizing is caused by for example chronic disease such as cancer or AIDS, treat high islets of langerhans mass formed by blood stasis and comprise islet cells disease, auxiliary treatment is brought out ovulation, stimulate thymus development, prevent the thymus function decline relevant with the age, the patient of treatment compromised immune, improve muscle strength, improve the mobility, old people's skin thickness is increased, metabolic balance, keep the kidney balance, stimulate osteoblast, osteanagenesis and regenerating bone or cartilage.
In addition, pharmaceutical composition of the present invention can be effective to promote growth of animal, increase animal to give milk in animal and produce hair with the disease relevant with the age of the immune system of producing hair, activation pet animals, treatment pet animals, the growth that promotes livestock animals and promotion sheep.
Pharmaceutical composition of the present invention can be with the form of aqueous solution according to various types of administering mode administrations.For example, pharmaceutical composition of the present invention is with for example intravenous injection of form administration, subcutaneous injection, intramuscular injection or the intravenous drip of injection solution.In addition, pharmaceutical composition of the present invention can with the administration of non-intestinal mode for example nose administration, through lung administration, percutaneous dosing or mucosal.In addition, pharmaceutical composition of the present invention can carry out the intestinal external administration with collyrium or the capsular form of filling solution.
Embodiment:
Describe the stability of ghrelin of the present invention in more detail by following test and embodiment, but should notice that the present invention also is subjected to the restriction of these tests and embodiment never in any form.
Unless indicate in addition, use following symbol and test method and instrument in the following description with specific meanings.
[symbol]
Dha: dehydroalanine
TFA: trifluoroacetic acid
HBTU:2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethylurea-hexafluorophosphate
The HOBt:1-hydroxybenzotriazole
TIPS: three-isopropyl silane
DIPEA: diisopropylethylamine
Fmoc: fluorenylmethyloxycarbonyl
Boc: tertbutyloxycarbonyl
TBu: the tert-butyl group
Trt: trityl
The DMAP:4-dimethyl aminopyridine
EDC:1-ethyl-3-(3-dimethylaminopropyl) carbodiimide
Pmc:2,2,5,7,8-pentamethyl benzo dihydropyran-6-sulfonyl
[device]
(A) automatic peptide synthesizer
Used biosystem: automatic 433A peptide synthesizer
(B) analyze the HPLC system that uses
Instrument: Tianjin, island LC-10A system
Post: YMC-Pack PROTEIN-RP (4.6mm Φ * 150mm) or
YMC-Pack?ODS-AM(4.6mmΦ×250mm)
Column temperature: 40 ℃
Eluent: in acetonitrile 0.1%TFA, the linear gradient of the highest 50% concentration
Flow velocity: 1mL/min
Detector: UV (210nm)
Admission space: 10-500 μ L
(C) five equilibrium HPLC system
Instrument: Waters 600 Multisolvent Delivery System
Post: YMC-Pack ODS-A (20mm Φ * 250mm) or
YMC-Pack?PROTEIN-RP(20mmΦ×250mm)
Eluent: the acetonitrile in 0.1%TFA or 5% acetic acid, linear gradient
Flow velocity: 10mL/min
Detector: UV (210 and 260nm)
Admission space: 1-2mL (, loading) with pump greater than 2mL
(D) storeroom
The LH-30 of thermostatic constant wet chamber
(Nagano?Kagaku)5℃/40℃
The LH-20 of prefabrication type thermostatic constant wet chamber
(Nagano?Kagaku)25℃
(E) mass spectrum
Instrument: Finnigan MAT TSQ700
Particle source: ESI
Detect the particle mode: positive
Spray voltage: 4.5kV
Capillary temperature: 250 ℃
Mobile phase: the water/methanol of 0.2% acetic acid (1/1) solution
Flow velocity: 0.2mL/min
Scanning area: m/z 300-1500
(F) analysis of aminoacid sequence
Instrument: Applied Biosystem 477A Sequencer (Perkin-Elmyer)
(G) analysis of aminoacid composition
Instrument: amino-acid analyzer L-8500 (Hitachi)
Sample: with the 6M-HCl that contains 0.1% phenol hydrolysis 24 hours in 110 ℃ airtight test tube.
Reference example: Synthesizing of the plain release peptide of human growth
Use the automatic peptide synthesizer, by Fmoc-Arg (Pmc)-HMP-resin (AppliedBiosystems Japan; 472mg, 0.25mmol) remove Fmoc repeatedly and insert synthetic Boc-Gly-Ser (tBu)-Ser (Trt)-Phe-Leu-Ser (tBu)-Pro-Glu (OtBu)-His (Boc)-Gln (Trt)-Arg (Pmc)-Val-Gln (Trt)-Gln (Trt)-Arg (Pmc)-Lys (Boc)-Glu (OtBu)-Ser (tBu)-Lys (Boc)-Lys (Boc)-Pro-Pro-Ala-Lys (Boc)-Leu-Gln (Trt)-Pro-Arg (the Pmc)-HMP resin that obtains of Fmoc-aminoacid (condition is that the N-terminal of glycine uses the Boc-glycine) by HBTU/HOBt.Resulting protected peptide-resin (1.7g) was handled 30 minutes with 1%TFA/5%TIPS/ dichloromethane solution (15mL).Collect peptide resin for several times, use the washing of 1%DIPEA (30mL) and dichloromethane (30mL) more successively with dichloromethane (30mL) washing.Resulting peptide-resin of removing Trt is expanded in N-Methyl pyrrolidone (10mL), with mixture and sad (144.2mg, 1.0mmol) and EDC-HCl (192mg, 1.0mmol) (0.25mmol) there are reaction down 16 hours in 31mg at DNAP.Filter and collect resulting resin, use N-Methyl pyrrolidone and washed with dichloromethane respectively, vacuum drying obtains protected peptide resin, is replaced by caprylyl on the 3-position of its serine side chain.In this resin, add then by 88%TFA/5% phenol/2%TIPS/5%H 2The deprotecting regent that O (15mL) forms at room temperature stirred the mixture 2 hours.Remove by filter resin, concentrated filtrate.Resulting residue is obtained precipitation with the ether processing, filter collecting precipitation.To precipitate drying and obtain the thick polypeptide of 900mg.The resulting thick polypeptide of 200mg is dissolved in the 10mL water, and it (is 60 minutes (flow velocitys: 10mL/min) of acetonitrile eluting of 0-60% with 5%TFA and linear gradient among the 20mm Φ * 250mm) that solution is added to YMC-Pack ODS-A post.Collect target eluting part and lyophilizing and obtain 60mg target peptide (the plain release peptide acetate of human growth: acetic acid content: 10.9%).
ESI-MS:3371 (value of calculation: 3370.9)
The Leu standard amino acid is formed: Ser; 3.43 (4), Glx; 5.93 (6), Gly; 1.01 (1), Ala; 1.00 (1), Val; 0.98 (1), Leu; 2 (2), Phe; 1.00 (1), Lys; 4.02 (4), His; 1.00 (1), Arg; 2.98 (3), Pro; (4) 3.93 (in the bracket is theoretical volume).
Amino acid sequence analysis: identify resulting polypeptide with human ghrelin (caprylyl-Ser on 3 is not protected).
Use step same as described above to obtain rat or other ghrelin.
In the following embodiments, use the ghrelin of reference example preparation.
Embodiment 1: The structural analysis of the catabolite of ghrelin
In order to guarantee the safety of ghrelin in aqueous solution, be necessary to understand the degradation reaction of ghrelin in aqueous solution.Therefore, thus by utilize a kind of ghrelin one by one the plain release peptide of human growth the catabolite of ghrelin is carried out the degradation process of structural analysis assessment ghrelin.
By the plain release peptide of about 5.0 μ mol (17mg) human growth is dissolved in Britton-Robinson buffer solution (pH 7.0: regulate with 0.04M phosphate/acetic acid/boric acid solution) and the 0.2M sodium hydrate aqueous solution, obtain containing the plain release peptide aqueous solution of human growth of about 0.15 μ mol/mL (0.5mg/mL).Resulting solution is loaded in the amber glass ampoule, uses fiery sealed ampoule.Store each ampoule 4 days and 14 days down at 40 ± 1 ℃.Store the back by the catabolite in the HPLC method detection aqueous solution, its result is as Fig. 1 (a) with (b).Shown in Fig. 1 (a), be to observe 2 main peaks (catabolite B and catabolite C) that are positioned at 24-28 minute in 40 ℃ of solution that storage obtained after 4 days down.Collect this two kinds of catabolite B and C, the catabolite to the plain release peptide of these human growth carries out structural analysis according to the methods below.
Catabolite B:
The ESI-MS analyser shows that this product quality is 3245, the plain release peptide of human growth (hereinafter being called " deacylation compound ") of the deacylation that this obtains with caprylyl by the plain release peptide of hydrolytic rupture human growth identical in quality.In addition, according to the analysis result that aminoacid sequence and the aminoacid of catabolite B are formed, the theoretical volume of this aminoacid composition and aminoacid sequence and deacylation compound is identical.Therefore, this proves that this catabolite B is exactly the deacylation compound.
Catabolite C:
The ESI-MS analyser shows that this product quality is 3227, [3-dehydroalanine] human growth element release peptide (hereinafter being called " Dha chemical compound ") that this obtains with caprylyl by the plain release peptide of β-eliminations human growth identical in quality.In addition, by with catabolite C and excessive reacting through the neutral ethyl mercaptan aqueous solution of 0.05M sodium hydrate aqueous solution, analyze according to ESI-MS, aminoacid sequence and the aminoacid of this product are formed, confirm that the plain release peptide of this product and [3-ethyl cystein] human growth is identical.That is to say that the value of calculation that the mass number 3289 that should be recorded by ESI-MS obtains with Dha chemical compound (3227) and ethyl mercaptan (62) are carried out addition is identical, and also detected ethyl cystein by aminoacid sequence and amino acid composition analysis.The plain release peptide of this product [3-ethyl cystein] human growth is obtained by Dha chemical compound and ethyl mercaptan generation necleophilic reaction.Therefore, this proves that this catabolite C is exactly the Dha chemical compound.
According to The above results, prove that as the catabolite in the neutral aqueous solution of the plain release peptide of the human growth of one of ghrelin be the deacylation compound that obtains of the caprylyl by the plain release peptide of hydrolytic scission human growth and the Dha chemical compound that obtained by the caprylyl of β-eliminations human growth element release peptide.
The HPLC result of the solution of storage after 14 days shown in Fig. 1 (b) except the peak of catabolite B and C, also observes other several peaks (catabolite D, E etc.).Therefore deacylation compound (catabolite B) and Dha chemical compound (catabolite C) are stored in the aqueous solution to investigate the mechanism of degradation of these catabolites.
According to method same as described above, deacylation compound or Dha compound dissolution are obtained containing the deacylation compound of about 0.15 μ mol/mL (0.5mg/mL) or the aqueous solution of Dha chemical compound in Britton-Robinson buffer solution (pH 7.0).Resulting solution is loaded in the amber glass ampoule, and ampoule seals with fire.The ampoule that the deacylation compound is housed stores 14 days down at 40 ± 1 ℃, and the ampoule that the Dha chemical compound is housed stores 3 days down at 40 ± 1 ℃.By the catabolite in the HPLC method detection aqueous solution, its result is as Fig. 1 (c) with (d) after storing.
Shown in Fig. 1 (c), in containing deacylation compound and the solution after 40 ℃ down store 14 days, observe an identical main peak of retention time (26 minutes) with the middle catabolite D of Fig. 1 (b).Collect the catabolite of this peak position and carry out ESI-MS, aminoacid sequence and amino acid composition analysis.According to these analysis results, this product is confirmed to be the plain release peptide (3-28) of deacylated tRNA base human growth.
ESI-MS:3101 (value of calculation: 3100.5).
In addition, shown in Fig. 1 (d), be to observe in 40 ℃ of solution that contain the Dha chemical compound that store down after 3 days one with Fig. 1 (b) in the identical big peak of retention time (36 minutes) of catabolite E.Collect the catabolite of this peak position and carry out ESI-MS, aminoacid sequence and amino acid composition analysis.According to these analysis results, this product is inferred to be [N a-CO-C (=CH 2)-OH] the plain release peptide (4-28) (hereinafter being called " the secondary catabolite of Dha chemical compound ") of human growth.
ESI-MS:3083 (value of calculation: 3083.5),
Aminoacid is formed: form consistent with the aminoacid of being inferred.
Aminoacid sequence: reactionless to first amino acid residue.
In addition, in Fig. 1 (d), also observe small peak about several 30-35 of being positioned at minutes.In Fig. 1 (b), also observe these peaks, infer that these catabolites are produced through the Dha chemical compound by the plain release peptide of human growth.
According to The above results; the degradation process that proof contains the aqueous solution of the plain release peptide of human growth is such: begin most to produce deacylation compound or Dha chemical compound by the plain release peptide of human growth; then respectively the cracking by this deacylation compound produce the plain release peptide (3-28) of deacylated tRNA base human growth and by more respectively cracking produce the plain release peptide (3-28) of deacylated tRNA base human growth, the cracking of this deacylation compound produces and other product of the secondary catabolite of Dha chemical compound.
Therefore, The above results proves in order to guarantee the stability of ghrelin in aqueous solution, need prevent from the hydrophobic group position as the feature structure of ghrelin various types of lytic responses to take place.
Embodiment 2: The stability of ghrelin in having the buffer solution of different pH value is (steady Qualitative test 1)
Use a kind of ghrelin---the plain release peptide evaluation of human growth contains the influence of the pH value of ghrelin solution.
The concentration of the plain release peptide of human growth with about 0.15 μ mol/mL (0.5mg/mL) is dissolved in the following aqueous solution.
0.1M HCl aqueous solution (pH:1.1)
Mcllvain buffer solution (pH:2.0,3.0,4.0,5.0,6.0,7.0,8.0)
Regulate pH with 0.1M aqueous citric acid solution and 0.2M sodium hydrogen phosphate aqueous solution.
Every kind of solution is stored 8,24,48 and 72 hours respectively under 25 ± 2 ℃, compare by this resulting solution of HPLC analyzing and testing and storage solution before.Calculate the peak area ratio that the plain release peptide of human growth, deacylation compound and Dha chemical compound account for the gross area.Find that pH value of solution is before storing and do not have significant change afterwards.The result is as shown in table 2.
Table 2:
Observed pH Account for the peak area ratio (%) of total peak area
The plain release peptide of human growth a) The deacylation compound b) The Dha chemical compound b)
pH?1 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ??1.1 ??1.2 ??1.2 ??1.2 ??1.2 ??98.89 ? 94.40? 86.00? 74.62? 65.04 ??0.27 ? 4.88? 13.31? 24.63? 34.17 ?0.21 ?0.15 ?0.09 ?0.04 ?0.00
pH?2 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ??2.0 ??2.0 ??2.0 ??2.0 ??2.0 ??99.07 ??98.82 ??98.24 ? 97.35? 96.46 ??0.10 ??0.34 ??0.89 ? 1.76? 2.59 ?0.20 ?0.20 ?0.23 ?0.24 ?0.25
pH?3 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ??3.0 ??3.1 ??3.1 ??3.1 ??3.1 ??99.07 ??98.98 ??98.77 ??98.40 ??98.03 ??0.10 ??0.20 ??0.38 ??0.67 ??0.98 ?0.20 ?0.20 ?0.23 ?0.24 ?0.25
pH?4 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ??4.0 ??4.0 ??4.0 ??4.0 ??4.0 ??99.07 ??99.07 ??98.92 ??98.69 ??98.44 ??0.09 ??0.12 ??0.21 ??0.37 ??0.51 ?0.20 ?0.20 ?0.21 ?0.22 ?0.23
pH?5 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ??5.0 ??5.0 ??5.0 ??5.0 ??5.0 ??99.06 ??99.04 ??98.91 ??98.59 ??98.37 ??0.10 ??0.14 ??0.24 ??0.45 ??0.61 ?0.21 ?0.21 ?0.21 ?0.23 ?0.24
pH?6 Before after the 8hr. after the 24hr. ??6.0 ??6.0 ??6.0 ??99.07 ??98.87 ??98.28 ??0.10 ??0.28 ??0.70 ?0.20 ?0.21 ?0.26
48hr. afterwards after the 72hr. ????6.0 ????6.0 ??? 97.64??? 96.92 ??? 1.33??? 1.97 ????0.31 ????0.35
?pH?7 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ????7.0 ????7.0 ????7.0 ????7.0 ????7.0 ????98.97 ????98.15 ??? 96.18??? 93.44??? 90.75 ????0.16 ????0.92 ??? 2.70??? 5.22??? 7.67 ????0.22 ????0.25 ????0.43 ????0.61 ????0.79
?pH?8 Before after the 8hr. after the 24hr. after the 48hr. after the 72hr. ????8.0 ????7.9 ????7.9 ????7.9 ????7.9 ????98.61 ??? 94.73??? 87.32??? 76.95??? 68.30 ????0.47 ??? 3.92??? 10.57??? 19.54??? 27.26 ????0.24 ????0.68 ??? 1.39??? 2.39??? 3.12
A) leukorrhagia line be that the human growth element release peptide peak area ratio of this position is less than 98%.
B) leukorrhagia line is that the peak area ratio of the deacylation compound of this position or Dha chemical compound is less than 1%.
Shown in above-mentioned table 2, the plain release peptide aqueous solution of human growth pH be 1.0 and within the short time, for example stored 8 hours at 8.0 o'clock after just produced deacylation compound above 3%.In addition, the plain release peptide aqueous solution of human growth pH be storing 24 hours at 7.0 o'clock after and pH be 2.0 and storing 48 hours at 6.0 o'clock after produced deacylation compound above 1%, and the ratio of human growth element release peptide is less than 98%.On the contrary, the pH scope is the generation that the solution of 3.0-5.0 has but suppressed this deacylation compound and Dha chemical compound.
Should be appreciated that the pH scope is the generation that solution that 2-7 is preferably 3-6 is suitable for suppressing deacylation compound and Dha chemical compound.Therefore, this proof is in order to guarantee the stability of ghrelin in aqueous solution, and the pH that needs regulator solution is that the preferred 3-6 of 2-7 is to prevent on as the hydrophobic group position of ghrelin feature structure various types of lytic responses taking place.
Embodiment 3: The stability of ghrelin in having the buffer solution of different pH value is (steady Qualitative test 2)
Use the buffer solution different, the stability of ghrelin in having the buffer solution of different pH value is tested with the buffer solution of embodiment 2.
With a kind of ghrelin---the plain release peptide of human growth is dissolved in pH and obtains containing the aqueous solution that the plain release peptide concentration of human growth is approximately 0.15 μ mol/mL (0.5mg/mL) in 2.1,3.1,4.0,5.0,6.0,7.0 and 7.9 the Britton-Robinson buffer solution, wherein by uniting the pH that the 0.04M phosphoric acid-acetic acid-boric acid aqueous solution that uses suitable proportion and 0.2M sodium hydrate aqueous solution are regulated this Britton-Robinson buffer solution.Resulting solution is loaded in the amber glass ampoule also with the fire sealing.For there are degree in computational dynamics constant and catabolite in the aqueous solution of ghrelin, ampoule is stored under 40 ± 1 ℃ the strict condition of storage.Utilize the concentration (remaining ratio) of the plain release peptide of measure of time human growth by HPLC.Also measure the pH of solution, find that significant change does not appear in the pH of solution.
Go out the kinetic constant of every kind of pH solution according to the continuous change calculations of remaining ratio, the result as shown in Figure 2.
In addition, each solution with different pH 40 ℃ store 1 day down after the peak area of its deacylation compound and Dha chemical compound account for the gross area ratio as shown in Figure 3.
As shown in Figure 2, the pH scope that proves the solution that can stably contain the plain release peptide of human growth is 3-6.PH be the remaining ratio of the plain release peptide of 3.1,4.0,5.0 and 6.0 growth from solution greater than 87%, these are worth greater than desired value (85%).
In addition, as shown in Figure 3, the pH scope is the generation that the solution of 3-6 can suppress the deacylation compound, can suppress the generation of Dha chemical compound and the pH scope is the solution of 2-7.
Can understand like this according to The above results, under the situation of using Britton-Robinson buffer solution, the pH scope is the generation that solution that 2-7 is preferably 3-6 relatively is suitable for suppressing deacylation compound and Dha chemical compound.Therefore, this proof is in order to guarantee the stability of ghrelin in aqueous solution, and the pH that needs regulator solution is that the preferred 3-6 of 2-7 is to prevent on as the hydrophobic group position of ghrelin feature structure various types of lytic responses taking place.
Embodiment 4: The kind of buffer solution is for the influence (test 1) of ghrelin stability
Use the stability of the plain release peptide of citrate buffer solution test vector generation for testing IC.
With a kind of ghrelin---it is that obtaining containing the plain release peptide concentration of human growth is the aqueous solution of about 0.15 μ mol/mL (0.5mg/mL) in 3.6,4.0,4.5 and 5.0 the citrate buffer solution that the plain release peptide of human growth is dissolved in pH.Resulting solution is loaded in the amber glass ampoule, and ampoule stored for 2 weeks with each ampoule down at 40 ± 1 ℃ then with the fire sealing.Store after 2 weeks, solution is carried out HPLC analyze, calculate remaining ratio according to the densitometer of the plain release peptide of human growth.Variation to pH value also detects.
The above results is summarised in the following table 3.
Table 3:
The pH of buffer solution After the preparation After 40 ℃ stored for 2 weeks down
?????3.5 Remaining ratio (%) ????100 ??????89
PH (observing) ????3.6 ??????3.5
?????4.0 Remaining ratio (%) ????100 ??????88
PH (observing) ????4.0 ??????4.0
?????4.5 Remaining ratio (%) ????100 ??????86
PH (observing) ????4.5 ??????4.4
?????5.0 Remaining ratio (%) ????100 ??????85
PH (observing) ????5.0 ??????4.9
As shown in table 3, if the plain release peptide of human growth is dissolved in the citrate buffer solution that the pH scope is 3.5-5.0, then the remaining ratio of the plain release peptide of human growth is 85-89%, and this is worth greater than desired value (85%).In addition, before storing and observe pH after storing and do not have significant change.Therefore, this proof can guarantee the stability of ghrelin in citrate buffer solution in the pH of 3.5-5.0 scope.
Embodiment 5: The buffer solution kind is for the influence (test 2) of ghrelin stability
Use the stability of the plain release peptide of glycine hydrochloride salt buffer solution or acetate buffer solution test vector generation for testing IC.
With a kind of ghrelin---it is that 2.5,3.1,3.6,4.2,4.6 and 4.8 0.05M glycine hydrochloride salt buffer solution or pH are that obtaining containing the plain release peptide concentration of human growth is the aqueous solution of about 0.15 μ mol/mL (0.5mg/mL) in 3.1,3.5,4.0,4.5 and 5.0 the 0.05M acetate buffer solution that the plain release peptide of human growth is dissolved in pH.Resulting solution is loaded in the amber glass ampoule, and ampoule seals with fire, and each ampoule stored for 2 weeks down at 40 ± 1 ℃ then.Store after 2 weeks, solution is carried out HPLC analyze to calculate purity.Variation to pH value also detects, and proves that significant change does not appear in the pH of solution.
The plain release peptide of human growth is stored 2 all remaining ratios afterwards in having the various buffer solution of different pH be summarised among Fig. 4.
As shown in Figure 4, if the plain release peptide of human growth is dissolved in the glycine hydrochloride salt buffer solution or acetate buffer solution that the pH scope is 2.5-5.0, then the remaining ratio of the plain release peptide of human growth is 87-97%, and this is worth greater than desired value (85%).
Result according to embodiment 2 (stability in Mcllvaine buffer solution), embodiment 3 (stability in Britton-Robinson buffer solution), embodiment 4 (stability in citrate buffer solution) and embodiment 5 (stability in glycine hydrochloride or acetate buffer solution) proves, in order to guarantee that the kind stable and assurance buffer solution of ghrelin in aqueous solution do not influence the stability of ghrelin in aqueous solution, the pH of regulator solution is considerable.
Embodiment 6: The stability of ghrelin in having solution in different concentration
The stability of ghrelin in having solution in different concentration is tested.
With a kind of ghrelin---the plain release peptide of human growth is dissolved in the aqueous solution of the following five kinds of concentration that obtain containing the plain release peptide of human growth in the 0.05M glycine hydrochloride salt buffer solution (pH 3.5), and it can be used as medicine and uses.
0.03nmol/mL(0.1μg/mL)；0.3nmol/mL(1.0μg/mL)；3.0nmol/mL(10.0μg/mL)；0.3μmol/mL(1.0mg/mL)；3μmol/mL(10mg/mL)。
Store each solution 24 hours down at 25 ± 2 ℃, after the storage, each solution is carried out HPLC analyze, calculate remaining ratio according to the plain release peptide densitometer of human growth.Variation to pH value also detects, and proves that significant change does not appear in the pH of solution.
The remaining ratio of solution before storing is defined as 100%, and the pH value variation of the remaining ratio after each solution stores and each solution is summarized in the following table 4.
Table 4:
Ghrelin concentration After just having prepared After 40 ℃ stored for 2 weeks down
??0.03nmol/mL Remaining ratio (%) ??100 ?62
??pH ??3.4 ?3.5
??0.3nmol/mL Remaining ratio (%) ??100 ?98
??pH ??3.5 ?3.5
??3.0nmol/mL Remaining ratio (%) ??100 ?98
??pH ??3.5 ?3.6
??0.3μmol/mL Remaining ratio (%) ??100 ?101
??pH ??3.4 ?3.3
??3μmol/mL Remaining ratio (%) ??100 ?101
??pH ??3.4 ?3.3
As shown in table 4, solution with plain release peptide of high concentration human growth (0.3nmol/mL, 3.0nmol/mL, 0.3 μ mol/mL and 3 μ mol/mL) is still keeping the stability of the plain release peptide of human growth in aqueous solution after 25 ℃ store 24 hours down, do not observe the decline of remaining ratio and the change of pH value.
On the contrary, in the situation of the solution with plain release peptide of low concentration human growth (0.03nmol/mL), the remaining ratio after solution stores is 62%; Yet do not observe the significant change of pH value yet and on the HPLC analysis chart, do not occur catabolite for example deacylation compound or Dha yet.Therefore, the reduction of the remaining ratio of the plain release peptide of human growth may be to cause the plain release peptide content of human growth to reduce on the chamber wall owing to it is adsorbed in this proof solution.Therefore infer that the solution with plain release peptide of low concentration human growth (0.03nmol/mL) also can keep the stability of the plain release peptide of human growth in aqueous solution after 25 ℃ store 24 hours down.
In a word, this proves that the concentration that ghrelin can stably be present in after regulating pH is that about 0.03nmol/mL is to the buffer solution of about 3 μ mol/mL.
Embodiment 7: Stability (the test of ghrelin in having the aqueous solution of different pH value 1)
Using a kind of ghrelin---the plain release peptide of human growth is tested the stability of ghrelin in having the aqueous solution of different pH value.
The plain release peptide of human growth is dissolved in the concentration purification water with 0.03 μ mol/mL (0.1mg/mL).The pH of this solution is 4.7.Solution is divided into 4 parts, a preservation separately, its excess-three part is adjusted to its pH value pH1.8 (with the hydrochloric acid of 17mM), pH 3.9 (with the hydrochloric acid of 0.20mM) and pH 7.8 (with the sodium hydroxide of 0.24mM) respectively by independent adding hydrochloric acid or sodium hydrate aqueous solution.
These four parts of solution are stored 1 day or 3 days down at 25 ± 2 ℃, after storing each solution is carried out HPLC and analyze, calculate the peak area ratio that the plain release peptide of human growth, deacylation compound and Dha chemical compound account for total peak area.
The above results is summarized in the following table 5.
Table 5:
Account for the peak area ratio (%) of total peak area
The plain release peptide of human growth a) The deacylation compound b) The Dha chemical compound b)
??pH?1.8 After 1 day before after 3 days ????99.16 ??? 97.68??? 95.01 ????0.12 ??? 1.46??? 4.08 0.20 0.31 0.34
??pH?3.9 After 1 day before after 3 days ????99.21 ????99.19 ????99.14 ????0.07 ????0.09 ????0.14 0.20 0.20 0.19
??pH?4.7 After 1 day before after 3 days ????99.20 ????99.12 ????98.94 ????0.06 ????0.15 ????0.28 0.20 0.21 0.21
??pH?7.8 After 1 day before after 3 days ????98.85 ??? 94.78??? 87.84 ????0.33 ??? 3.65??? 9.84 0.24 0.86 1.65
A) leukorrhagia line be that the human growth element release peptide peak area ratio of this position is less than 98%.
B) leukorrhagia line is that the peak area ratio of the deacylation compound of this position or Dha chemical compound is less than 1%.
As shown in table 5, pH 1.8 and 7.8 aqueous solution have just produced after only storing 1 day and have surpassed 1% deacylation compound, and the ratio of human growth element release peptide is lower than 98%.In addition, the generation ratio of pH 1.8 and 7.8 aqueous solution its Dha chemical compound after storing 3 days is above 1%.On the contrary, in the aqueous solution of pH 3.9 and 4.7, but suppressed the generation of deacylation compound and Dha chemical compound.
Therefore, should be readily appreciated that having the pH scope is the generation that the aqueous solution of 2-7 relatively is suitable for suppressing deacylation compound and Dha chemical compound.Therefore, this proof is in order to guarantee the stability of ghrelin in aqueous solution, and the pH that needs regulator solution is that 2-7 is to prevent on as the hydrophobic group position of ghrelin feature structure various types of lytic responses taking place.
Embodiment 8: Stability (the test of ghrelin in having the aqueous solution of different pH value 2)
Using a kind of ghrelin---plain release peptide (1-7) amide of human growth is tested the stability of ghrelin in having the aqueous solution of different pH value.
Plain release peptide (1-7) amide of human growth is the common aminoacid sequence of the ghrelin that obtained by mammal, bird or Fish (referring to table 1), and it has the biological activity identical with the plain release peptide of human growth (the open WO 01/07475 of international monopoly).
Plain release peptide (1-7) amide of human growth is dissolved in the concentration purification water with about 0.12 μ mol/mL (0.1mg/mL).The pH of this solution is 5.0.Solution is divided into 4 parts, a preservation separately, its excess-three part is adjusted to its pH value pH 1.8 (with the hydrochloric acid of 17mM), pH 4.1 (with the hydrochloric acid of 0.05mM) and pH 7.9 (with the sodium hydroxide of 0.20mM) respectively by independent adding hydrochloric acid or sodium hydrate aqueous solution.
These four parts of solution are stored 1 day or 3 days down at 25 ± 2 ℃; after storing each solution is carried out HPLC and analyze, calculate the peak area ratio that plain release peptide (1-7) amide of human growth, plain release peptide (1-7) amide of deacylated tRNA base human growth and plain release peptide (1-7) amide of [3-dehydroalanine]-human growth account for total peak area.
The above results is summarized in the following table 6.
Table 6:
Peak area accounts for the ratio (%) of total peak area
Plain release peptide (1-7) amide of human growth a) Plain release peptide (1-7) amide of deacylated tRNA base human growth b) Plain release peptide (1-7) amide of [3-dehydroalanine] human growth b)
??pH?1.8 After 1 day before after 3 days ??98.39 ? 95.40? 90.11 ??0.98 ? 3.75? 8.89 ????0.02 ????0.07 ????0.10
??pH?4.1 After 1 day before after 3 days ??98.76 ??98.68 ??98.60 ??0.72 ??0.73 ??0.77 ????0.00 ????0.03 ????0.01
??pH?5.0 After 1 day before after 3 days ??98.69 ??98.57 ??98.51 ??0.70 ??0.73 ??0.79 ????0.00 ????0.02 ????0.06
??pH?7.9 After 1 day before after 3 days ??98.22 ? 95.89? 92.54 ? 1.12? 3.21? 6.21 ????0.03 ????0.23 ????0.54
A) leukorrhagia line be that human growth element release peptide (1-7) the amide peak area ratio of this position is less than 98%.
B) leukorrhagia line be that the peak area ratio of plain release peptide (1-7) amide of human growth of this position or [3-dehydroalanine]-human growth element release peptide (1-7) amide is less than 1%.
As shown in table 6, the ratio that plain release peptide (1-7) amide of human growth only stored in the aqueous solution of pH 1.8 and 7.8 after 1 day just is lower than 98%.On the contrary, in the aqueous solution of pH 4.1 and 5.0, but suppressed the generation of catabolite.
Therefore, should be readily appreciated that having the pH scope is that the aqueous solution of 2-7 is to contain a kind of ghrelin---the relatively more suitable aqueous solution of plain release peptide (1-7) amide of human growth.Therefore, this proof is in order to guarantee the stability of ghrelin in aqueous solution, and the pH that needs regulator solution is that 2-7 is to prevent on as the hydrophobic group position of ghrelin feature structure various types of lytic responses taking place.
Embodiment 9: The inhibition of anti-adsorbent in the aqueous solution that contains the low concentration ghrelin inhaled Attached effect
In embodiment 6, but show that this ghrelin can be adsorbed on the chamber wall in the solution of the ghrelin with hyoscine concentration.Therefore resisting the absorption inhibition effect of adsorbent in containing the solution of ghrelin tests.
Can the option table surface-active agent just the polyoxyethylene sorbitol monoleate (hereinafter be called tween 80) and benzalkonium chloride as anti-adsorbent.
The concentration of the plain release peptide of human growth with 0.3nmol/mL (1.0 μ g/mL) is dissolved in 5% mannitol-0.05M glycine hydrochloride salt buffer solution (pH 3.5), but this concentration is the concentration of ghrelin hyoscine.Concentration with 0.01% or 0.1% adds anti-adsorbent in solution.
Measure the plain release peptide concentration of human growth of each solution after the preparation immediately by the HPLC method, then above-mentioned solution is placed teat glass.Afterwards, again each solution is placed a new teat glass, repeat same operation 5 and 10 times, measure the plain release peptide concentration of human growth in the solution of various treated mistakes by the HPLC method.In contrast, the solution that does not contain anti-adsorbent is tested.
The test tube that use is made by polypropylene replaces teat glass to repeat same step.
The above results is summarized in the table 7.
Table 7:
Anti-adsorbent Initial concentration Teat glass a) The polypropylene test tube b)
Kind Concentration 5 times 10 times 5 times 10 times
Do not have ?- ??100 ????0 ????0 ????51 ????19
Tween 80 ?0.01％ ??100 ????17 ????0 ????90 ????90
?0.1％ ??100 ????20 ????5 ????93 ????91
Benzalkonium chloride ?0.01％ ??100 ????81 ????64 ????97 ????96
?0.1％ ??100 ????99 ????99 ????99 ????101
A) Asahi Techno-glass company: 10mL
B) CORNING company: 15mL
Table 7 clearlys show, solution is no matter be wherein all obviously reductions of the plain release peptide concentration of human growth after teat glass or the displacement of polypropylene test tube.Especially, the plain release peptide concentration of the human growth of this solution is reduced to the degree that can not be gone out by the HPLC analyzing and testing in the metathetical situation with teat glass.
On the contrary, if in solution, add tween with 0.01% or 0.1% concentration 80 as anti-adsorbent, can observe this solution the polypropylene test tube is had high absorption inhibition effect, if the concentration with 0.01% or 0.1% adds benzalkonium chloride as anti-adsorbent in solution in addition, also can observe this solution all has high inhibition absorption to teat glass and polypropylene test tube effect.
Therefore should be readily appreciated that, but for the solution of the ghrelin that contains hyoscine concentration, tween 80 and benzalkonium chloride can suppress this ghrelin effectively and adsorbed by chamber wall.Therefore, this proves that anti-adsorbent can prevent effectively that ghrelin is adsorbed in preparation, long term storage and administration process.
Embodiment 10: The absorption of saccharide in the aqueous solution that contains the low concentration ghrelin suppresses to imitate Really
In embodiment 9, show by using anti-adsorbent can suppress the absorption of ghrelin on the chamber wall of solution.In the present embodiment the absorption inhibition effect of saccharide is tested.
Use 5% Osmitrol as sugar aqueous solution.Concentration with about 3nmol/mL (10 μ g/mL) and about 30nmol/mL (100 μ g/mL) is dissolved in the plain release peptide of human growth in 5% Osmitrol, but this concentration is the concentration of ghrelin hyoscine.
Measure the plain release peptide concentration of human growth of each solution after the preparation immediately by the HPLC method, then above-mentioned solution is placed the polypropylene test tube.Afterwards, again each solution is placed a new polypropylene test tube, repeat same operation altogether 5 times, measure the plain release peptide concentration of human growth in the solution of various treated mistakes by the HPLC method.In contrast, test containing the normal saline solution that the plain release peptide concentration of human growth is approximately 3nmol/mL (10 μ g/mL) and about 30nmol/mL (100 μ g/mL).
The above results is summarized in the table 8.
Table 8:
Anti-adsorbent Initial concentration The polypropylene test tube b)
Kind The plain release peptide concentration of human growth 5 times
Normal saline ??10μg/mL ??100 ??32
??100μg/mL ??100 ??89
5% Osmitrol ??10μg/mL ??100 ??98
??100μg/mL ??100 ??99
B) CORNING company: 15mL
Table 8 clearlys show that the plain release peptide concentration of its human growth significantly reduces after with the polypropylene test tube normal saline solution being shifted.Especially, in containing the normal saline solution that concentration is 10 μ g/mL, the plain release peptide concentration of the human growth of this solution is reduced to 32%.
On the contrary, in containing 5% Osmitrol that concentration is 10 μ g/mL and 100 μ g/mL, all observe above-mentioned solution the polypropylene test tube is all had high absorption inhibition effect.
Therefore should be readily appreciated that, but for the solution of the ghrelin that contains hyoscine concentration, saccharide can suppress this ghrelin effectively and be adsorbed by chamber wall.Therefore, this proof saccharide can prevent effectively that ghrelin is adsorbed in preparation, long term storage and administration process.
Embodiment 11: The preparation of the on-the-spot preparation of solution and stability thereof
As the on-the-spot preparation of pharmaceutical composition of the present invention, prepared freeze-dried powder below and the dissolubility of this powder has been estimated.
The concentration of the plain release peptide of human growth with about 0.3nmol/mL (1.0 μ g/mL) is dissolved in 5% mannitol-0.05M glycine hydrochloride salt buffer solution (pH 3.5), measure the plain release peptide concentration of human growth of this solution after the preparation immediately by the HPLC method, also measured the pH value of this solution.Consequently the plain release peptide concentration of this human growth is 1.0 μ g/mL, and pH is 3.5.Then ,-25 ℃ of following vacuum freeze-dryings 24 hours, resulting powder was further 20 ℃ of following vacuum dryings 24 hours with this solution.Resulting freeze-dried powder is the white solid with good appearance.
Then, this freeze-dried powder is dissolved in purifies waste water, its consumption is identical with the amount that lyophilizing begins to reduce, the dissolubility of freeze-dried powder and the pH of resultant solution are detected.
Consequently the dissolubility of this freeze-dried powder is fairly good, looks not have insoluble matter in the solution, and the dissolubility of visible this powder relatively is suitable for the on-the-spot preparation of solution.The plain release peptide concentration of this human growth is 1.0 μ g/mL, its with after solution has just prepared or the plain release peptide concentration of human growth before the lyophilizing identical, that is to say the not reduction of content of observing said preparation after lyophilizing.The pH value of solution is 3.5 in addition, after this value and solution have just prepared or the pH value before the lyophilizing identical.The aqueous solution that therefore, can stably be contained the plain release peptide of human growth by freeze-dried powder of the present invention.
Therefore, the solution that contains ghrelin of pH through overregulating that uses solution can obtain containing the freeze-dried powder of ghrelin through lyophilization, and resulting freeze drying powder preparations can be used as the on-the-spot preparation of pharmaceutical composition of the present invention.
Preparation embodiment 1: Preparation contains the medicine that pH is conditioned the plain release peptide of human growth afterwards Compositions
With a kind of ghrelin---obtaining containing the plain release peptide concentration of human growth during the plain release peptide of human growth is dissolved in and purifies waste water is the aqueous solution of about 0.15 μ mol/mL (0.5mg/mL).Regulate the pH to 4.0 of this solution by adding the 0.1M hydrochloric acid solution, obtain the pharmaceutical composition that contains the plain release peptide of human growth of pharmaceutical solutions form.
Preparation embodiment 2: Preparation contains the glycine hydrochloride salt buffer solution of rat ghrelin Pharmaceutical composition
With a kind of ghrelin---the rat ghrelin is dissolved in the 0.05M glycine hydrochloride salt buffer solution (pH 3.5) with the concentration that is approximately 0.15 μ mol/mL (0.5mg/mL), obtains the pharmaceutical composition that contains the rat ghrelin of pharmaceutical solutions form.The pH of this solution is 3.5.
Preparation embodiment 3: Preparation contains the sweet ammonia of the plain release peptide of [3-serine (acetyl group)] human growth The pharmaceutical composition of acid hydrochloride buffer solution
With a kind of ghrelin---the plain release peptide of [3-serine (acetyl group)] human growth is dissolved in the 0.05M glycine hydrochloride salt buffer solution (pH 3.5) with the concentration that is approximately 0.15 μ mol/mL (0.1mg/mL), obtains the pharmaceutical composition that contains the plain release peptide of [3-serine (acetyl group)] human growth of pharmaceutical solutions form.The pH of this solution is 3.5.
Preparation embodiment 4: Preparation contains the plain release peptide of [3-serine (phenyl propiono)] human growth The pharmaceutical composition of glycine hydrochloride salt buffer solution
With a kind of ghrelin---the plain release peptide of [3-serine (phenyl propiono)] human growth is dissolved in the 0.05M glycine hydrochloride salt buffer solution (pH 3.5) with the concentration that is approximately 0.15 μ mol/mL (0.5mg/mL), obtains the pharmaceutical composition that contains the plain release peptide of [3-serine (phenyl propiono)] human growth of pharmaceutical solutions form.The pH of this solution is 3.5.
Preparation embodiment 3: Preparation contains the glycine hydrochloride of plain release peptide (1-5) amide of human growth The pharmaceutical composition of buffer solution
With a kind of ghrelin---plain release peptide (1-5) amide of human growth is dissolved in the 0.05M glycine hydrochloride salt buffer solution (pH 3.5) with the concentration that is approximately 0.15 μ mol/mL (0.5mg/mL), obtains the pharmaceutical composition that contains the plain release peptide of [3-serine (phenyl propiono)] human growth of pharmaceutical solutions form.The pH of this solution is 3.5.
Industrial usability
As mentioned above, the invention provides the pharmaceutical composition that can stably contain ghrelin, and this preparation of the present invention can prevent that ghrelin from being adsorbed by chamber wall. Therefore, the invention provides the pharmaceutical composition that ghrelin can not reduce in preparation, long term storage or administration process.
Sequence table
<110>DAIICHI?SUNTORY?PHARMA?CO.，LTD.
＜120〉contain the pharmaceutical composition of ghrelin
<130>DSP-78
<150>JP?2002-146155
<151>2002-05-21
<160>21
＜170〉PatentIn is 3.1 editions
<210>1
<211>28
<212>PRT
＜213〉people
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the human endogenous peptide of somatropin secretin
<400>1
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Arg?Val?Gln?Gln?Arg?Lys
1????????????????5??????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20?????????????????25
<210>2
<211>27
<212>PRT
＜213〉people
<220>
＜221〉peptide
<222>(1)..(27)
＜223〉aminoacid sequence of the human endogenous peptide of somatropin secretin (27 aminoacid)
<400>2
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Arg?Val?Gln?Arg?Lys?Glu
1????????????????5??????????????????10??????????????????15
Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20?????????????????25
<210>3
<211>28
<212>PRT
＜213〉brown rat
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of rat of somatropin secretin
<400>3
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Ala?Gln?Gln?Arg?Lys
1???????????????5???????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20?????????????????25
<210>4
<211>27
<212>PRT
＜213〉brown rat
<220>
＜221〉peptide
<222>(1)..(27)
＜223〉aminoacid sequence of the endogenous peptide of rat of somatropin secretin
<400>4
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Ala?Gln?Arg?Lys?Glu
1???????????????5???????????????????10??????????????????15
Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20??????????????????25
<210>5
<211>28
<212>PRT
＜213〉house mouse
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of mice of somatropin secretin
<400>5
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Ala?Gln?Gln?Arg?Lys
1???????????????5???????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20??????????????????25
<210>6
<211>28
<212>PRT
＜213〉pig
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of pig of somatropin secretin
<400>6
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Val?Gln?Gln?Arg?Lys
1???????????????5???????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Ala?Ala?Lys?Leu?Lys?Pro?Arg
20??????????????????25
<210>7
<211>27
<212>PRT
＜213〉bull
<220>
＜221〉peptide
<222>(1)..(27)
＜223〉aminoacid sequence of the endogenous peptide of the cattle of somatropin secretin (27 aminoacid)
<400>7
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Leu?Gln?Arg?Lys?Glu
1???????????????5????????????????????10?????????????????15
Ala?Lys?Lys?Pro?Ser?Gly?Arg?Leu?Lys?Pro?Arg
20?????????????????25
<210>8
<211>27
<212>PRT
＜213〉sheep
<220>
＜221〉peptide
<222>(1)..(27)
＜223〉aminoacid sequence of the endogenous peptide of the sheep of somatropin secretin (27 aminoacid)
<400>8
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Leu?Gln?Arg?Lys?Glu
1???????????????5???????????????????10?????????????????15
Pro?Lys?Lys?Pro?Ser?Gly?Arg?Leu?Lys?Pro?Arg
20??????????????????25
<210>9
<211>28
<212>PRT
＜213〉tame Canis familiaris L.
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of Canis familiaris L. of somatropin secretin
<400>9
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?Gln?Lys?Leu?Gln?Gln?Arg?Lys
1????????????????5??????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Gln?Pro?Arg
20?????????????????25
<210>10
<211>21
<212>PRT
＜213〉Japanese Anguillar japonica
<220>
＜221〉peptide
<222>(1)..(21)
＜223〉aminoacid sequence of the endogenous peptide of Anguillar japonica of somatropin secretin.This peptide at the C-end by amidatioon.
<400>10
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Ser?Gln?Arg?Pro?Gln?Gly?Lys?Asp?Lys
1???????????????5???????????????????10??????????????????15
Lys?Pro?Pro?Arg?Val
20
<210>11
<211>23
<212>PRT
＜213〉rainbow trout
<220>
＜221〉peptide
<222>(1)..(23)
＜223〉aminoacid sequence of the endogenous peptide of the rainbow trout of somatropin secretin (23 aminoacid).This peptide at the C-end by amidatioon.
<400>11
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Ser?Gln?Lys?Pro?Gln?Val?Arg?Gln?Gly
1???????????????5???????????????????10??????????????????15
Lys?Gly?Lys?Pro?Pro?Arg?Val
20
<210>12
<211>20
<212>PRT
＜213〉rainbow trout
<220>
＜221〉peptide
<222>(1)..(20)
＜223〉aminoacid sequence of the endogenous peptide of the rainbow trout of somatropin secretin (20 aminoacid).This peptide at the C-end by amidatioon.
<400>12
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Ser?Gln?Lys?Pro?Gln?Gly?Lys?Gly?Lys
1???????????????5???????????????????10??????????????????15
Pro?Pro?Arg?Val
20
<210>13
<211>24
<212>PRT
＜213〉tame chicken
<220>
＜221〉peptide
<222>(1)..(24)
＜223〉aminoacid sequence of the endogenous peptide of tame chicken of somatropin secretin
<400>13
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Thr?Tyr?Lys?Asn?Ile?Gln?Gln?Gln?Lys
1???????????????5???????????????????10??????????????????15
Gly?Thr?Arg?Lys?Pro?Thr?Ala?Arg
20
<210>14
<211>24
<212>PRT
＜213〉tame chicken
<220>
＜221〉peptide
<222>(1)..(24)
＜223〉aminoacid sequence of the endogenous peptide of tame chicken of somatropin secretin
<400>14
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Thr?Tyr?Lys?Asn?Ile?Gln?Gln?Gln?Lys
1???????????????5???????????????????10??????????????????15
Asp?Thr?Arg?Lys?Pro?Thr?Ala?Arg
20
<210>15
<211>26
<212>PRT
＜213〉tame chicken
<220>
＜221〉peptide
<222>(1)..(26)
＜223〉aminoacid sequence of the endogenous peptide of tame chicken of somatropin secretin
<400>15
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Thr?Tyr?Lys?Asn?Ile?Gln?Gln?Gln?Lys
1???????????????5???????????????????10??????????????????15
Asp?Thr?Arg?Lys?Pro?Thr?Ala?Arg?Leu?His
20???????????????????25
<210>16
<211>27
<212>PRT
＜213〉frog
<220>
＜221〉peptide
<222>(1)..(27)
＜223〉aminoacid sequence of the endogenous peptide of the frog of somatropin secretin
<400>16
Gly?Leu?Thr?Phe?Leu?Ser?Pro?Ala?Asp?Met?Gln?Lys?Ile?Ala?Glu?Arg
1???????????????5???????????????????10?????????????????15
Gln?Ser?Gln?Asn?Lys?Leu?Arg?His?Gly?Asn?Met
20?????????????????25
<210>17
<211>28
<212>PRT
＜213〉frog
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of the frog of somatropin secretin
<400>17
Gly?Leu?Thr?Phe?Leu?Ser?Pro?Ala?Asp?Met?Gln?Lys?Ile?Ala?Glu?Arg
1???????????????5???????????????????10??????????????????15
Gln?Ser?Gln?Asn?Lys?Leu?Arg?His?Gly?Asn?Met?Asn
20??????????????????25
<210>18
<211>20
<212>PRT
＜213〉tilapia
<220>
＜221〉peptide
<222>(1)..(20)
＜223〉aminoacid sequence of the endogenous peptide of tilapia of somatropin secretin.Amidatioon
<400>18
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Ser?Gln?Lys?Pro?Gln?Asn?Lys?Val?Lys
1???????????????5???????????????????10??????????????????15
Ser?Ser?Arg?Ile
20
<210>19
<211>22
<212>PRT
＜213〉Silurus asotus fish
<220>
＜221〉peptide
<222>(1)..(22)
＜223〉aminoacid sequence of the endogenous peptide of Silurus asotus fish of somatropin secretin.This peptide at the C-end by amidatioon.
<400>19
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Thr?Gln?Lys?Pro?Gln?Asn?Arg?Gly?Asp
1???????????????5???????????????????10?????????????????15
Arg?Lys?Pro?Pro?Arg?Val
20
<210>20
<211>23
<212>PRT
＜213〉Silurus asotus fish
<220>
＜221〉peptide
<222>(1)..(23)
＜223〉aminoacid sequence of the endogenous peptide of Silurus asotus fish of somatropin secretin
<400>20
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Thr?Gln?Lys?Pro?Gln?Asn?Arg?Gly?Asp
1???????????????5???????????????????10??????????????????15
Arg?Lys?Pro?Pro?Arg?Val?Gly
20
<210>21
<211>28
<212>PRT
＜213〉horse
<220>
＜221〉peptide
<222>(1)..(28)
＜223〉aminoacid sequence of the endogenous peptide of horse of somatropin secretin.
<400>21
Gly?Ser?Ser?Phe?Leu?Ser?Pro?Glu?His?His?Lys?Val?Gln?His?Arg?Lys
1???????????????5???????????????????10??????????????????15
Glu?Ser?Lys?Lys?Pro?Pro?Ala?Lys?Leu?Lys?Pro?Arg
20?????????????????25

Claims (27)

1. the pharmaceutical composition that contains the ghrelin or derivatives thereof, the pH that wherein is dissolved with the aqueous solution of this ghrelin is 2-7.
2. according to the pharmaceutical composition of claim 1, wherein said pH is 3-6.
3. according to the pharmaceutical composition of claim 1 or 2, wherein also contain pH regulator agent or buffer agent.
4. according to the pharmaceutical composition of claim 3, wherein said pH regulator agent is selected from following one or more: hydrochloric acid, sulphuric acid, nitric acid, boric acid, carbonic acid, heavy carbonic, gluconic acid, sodium hydroxide, potassium hydroxide, ammonia, citric acid, monoethanolamine, lactic acid, acetic acid, succinic acid, fumaric acid, maleic acid, phosphoric acid, pyrovinic acid, malic acid, propanoic acid, trifluoroacetic acid and their salt.
5. according to the pharmaceutical composition of claim 3, wherein said buffer agent is selected from following one or more: glycine, acetic acid, citric acid, boric acid, phthalic acid, phosphoric acid, succinic acid, lactic acid, tartaric acid, carbonic acid, hydrochloric acid, sodium hydroxide and their salt.
6. according to pharmaceutical composition any among the claim 3-5, wherein said pH regulator agent or the concentration of buffer agent in solution are 0.01Mm-1000Mm.
7. according to pharmaceutical composition any among the claim 1-6, wherein said solution is buffer solution.
8. according to the pharmaceutical composition of claim 7, wherein said buffer solution is glycine hydrochloride salt buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer, phosphate-acetate-borate buffer solution or phthalate buffer.
9. according to pharmaceutical composition any among the claim 1-8, the concentration of wherein said ghrelin in solution is 0.03nmol/mL to 6 μ mol/mL.
10. according to pharmaceutical composition any among the claim 1-9, wherein said ghrelin is an acetate.
11. according to pharmaceutical composition any among the claim 1-10, wherein said ghrelin is the plain release peptide of human growth.
12., wherein also contain anti-adsorbent according to pharmaceutical composition any among the claim 1-11.
13. according to the pharmaceutical composition of claim 12, the concentration of wherein said anti-adsorbent is 0.001%-5%.
14. according to the pharmaceutical composition of claim 12 or 13, wherein said anti-adsorbent is a surfactant.
15. contain the pharmaceutical composition of ghrelin, wherein contain by any powder that described solution drying obtains among the claim 1-14.
16. according to the pharmaceutical composition of claim 15, wherein said powder is a freeze-dried powder.
17. prevent the hydrophobic group of ghrelin or derivatives thereof the method for degraded takes place in containing the solution of ghrelin, it comprises that the pH of regulator solution is 2-7.
18., wherein described pH value of solution is adjusted to 3-6 according to the method for claim 17.
19., wherein also comprise pH regulator agent or buffer agent according to the method for claim 17 or 18.
20. according to the method for claim 19, comprising being selected from one or more following pH regulator agent: hydrochloric acid, sulphuric acid, nitric acid, boric acid, carbonic acid, heavy carbonic, gluconic acid, sodium hydroxide, potassium hydroxide, ammonia, citric acid, monoethanolamine, lactic acid, acetic acid, succinic acid, fumaric acid, maleic acid, phosphoric acid, pyrovinic acid, malic acid, propanoic acid, trifluoroacetic acid and their salt.
21. according to the method for claim 19, comprising being selected from one or more following buffer agents: glycine, acetic acid, citric acid, boric acid, phthalic acid, phosphoric acid, succinic acid, lactic acid, tartaric acid, carbonic acid, hydrochloric acid, sodium hydroxide and their salt.
22. according to method any among the claim 19-21, wherein said pH regulator agent or the concentration of buffer agent in solution are 0.01Mm to 1000Mm.
23. according to method any among the claim 17-22, wherein said solution is buffer solution.
24. according to the method for claim 23, wherein said buffer solution is glycine hydrochloride salt buffer, acetate buffer, citrate buffer, lactate buffer, phosphate buffer, citric acid-phosphate buffer, phosphate-acetate-borate buffer solution or phthalate buffer.
25. according to method any among the claim 17-24, the concentration of wherein said ghrelin in solution is 0.03nmol/mL to 6 μ mol/mL.
26. according to method any among the claim 17-25, wherein said ghrelin is an acetate.
27. the method any according to claim 17-26, wherein said ghrelin are human growth element release peptides.
CNB038028301A 2002-05-21 2003-05-21 Pharmaceutical composition containing ghrelin Expired - Lifetime CN100411683C (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2002146155 2002-05-21
JP146155/2002 2002-05-21

Publications (2)

Publication Number Publication Date
CN1622822A true CN1622822A (en) 2005-06-01
CN100411683C CN100411683C (en) 2008-08-20

Family

ID=29545111

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB038028301A Expired - Lifetime CN100411683C (en) 2002-05-21 2003-05-21 Pharmaceutical composition containing ghrelin

Country Status (11)

Country Link
US (3) US20060166871A1 (en)
EP (1) EP1506786B1 (en)
JP (1) JP5000848B2 (en)
KR (1) KR20050012224A (en)
CN (1) CN100411683C (en)
AU (1) AU2003235401B2 (en)
BR (1) BR0306685A (en)
CA (1) CA2471879C (en)
ES (1) ES2615484T3 (en)
IL (2) IL162796A0 (en)
WO (1) WO2003097083A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101541341B (en) * 2006-09-27 2013-06-05 益普生制药两合公司 Analogs of ghrelin substituted at the N-terminal
CN114306225A (en) * 2019-05-07 2022-04-12 前沿生物药业(南京)股份有限公司 Stable esbociclib composition

Families Citing this family (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4562394B2 (en) 2001-12-18 2010-10-13 アリゼ、ファルマ、エスアーエス Pharmaceutical composition comprising non-acylated ghrelin and therapeutic use thereof
US7666833B2 (en) 2001-12-18 2010-02-23 Alizé Pharma SAS Pharmaceutical compositions comprising unacylated ghrelin and therapeutical uses thereof
CN100411683C (en) 2002-05-21 2008-08-20 阿斯比奥制药株式会社 Pharmaceutical composition containing ghrelin
CN1688696A (en) * 2002-09-18 2005-10-26 蒙特利尔大学医疗中心 GHRH analogs
WO2005039624A1 (en) * 2003-10-24 2005-05-06 Theratechnologies Inc. Use of ghrelin and unacylated ghrelin compostions in insulin-related disease conditions
CN1321131C (en) * 2003-12-26 2007-06-13 李宁 Pig Ghrelin derivative, its encoding gene and application
KR20070050871A (en) * 2004-05-11 2007-05-16 더 거번먼트 오브 더 유나이티드 스테이츠 오브 어메리카 애즈 레프리젠티드 바이 더 세크러터리 오브 더 디파트먼트 오브 헬쓰 앤드 휴먼 써비시즈 How to use ghrelin to inhibit proinflammatory cytokine expression
EP1781311A4 (en) * 2004-08-18 2010-02-17 Elixir Pharmaceuticals Inc Growth-hormone secretagogues
EP1800689B1 (en) 2004-08-24 2016-12-28 Daiichi Sankyo Company, Limited Liquid preparation of physiologically active peptide
CN101107013A (en) * 2004-11-22 2008-01-16 阿纳迪斯有限公司 Bioactive compositions
JPWO2006112443A1 (en) * 2005-04-15 2008-12-11 国立大学法人信州大学 Novel method of using triacetin and test aid for ultrasonic diagnosis
ATE501248T1 (en) * 2005-07-02 2011-03-15 Arecor Ltd STABLE AQUEOUS SYSTEMS WITH PROTEINS
CN101312745A (en) * 2005-11-21 2008-11-26 国立大学法人宫崎大学 Therapeutic agent for skin or skin repair-promoting agent comprising ghrelin, derivative thereof or substance capable of acting on ghs-r1a as active ingredient
US20090053334A1 (en) 2006-01-31 2009-02-26 Nat. Uni. Corp. Hokkaido University Ghrelin production promoter
US20100227806A1 (en) 2006-03-10 2010-09-09 Tulipano Giovanni Use Of A Ghrelin Agonist To Improve Catabolic Effects Of Glucocorticoid Treatment
NZ571183A (en) * 2006-03-13 2010-09-30 Liat Mintz Use of ghrelin splice variant for treating cachexia and/or anorexia and/or anorexia-cachexia and/or malnutrition and/or lipodystrophy and/or muscle wasting and/or appetite stimulation
US7763707B2 (en) 2006-03-13 2010-07-27 Liat Mintz Use of ghrelin splice variant for treating cachexia and/or anorexia and/or anorexia-cachexia and/or malnutrition and/or lipodystrophy and/or muscle wasting and/or appetite-stimulation
GB0700523D0 (en) * 2007-01-11 2007-02-21 Insense Ltd The Stabilisation Of Proteins
JP5485876B2 (en) 2007-05-31 2014-05-07 アリゼ ファーマ エスアーエス Unacylated ghrelin as a therapeutic agent in the treatment of metabolic diseases
US8318664B2 (en) 2007-05-31 2012-11-27 Alize Pharma Sas Unacylated ghrelin fragments as therapeutic agent in the treatment of obesity
JP2010538993A (en) * 2007-09-11 2010-12-16 モンドバイオテック ラボラトリーズ アクチエンゲゼルシャフト Use of the peptide Pro-Gly-Thr-Cys-Glu-Ile-Cys-Ala-Tyr-Ala-Ala-Cys-Thr-Gly-Cys as a therapeutic agent
EP2457590B1 (en) 2008-05-01 2014-12-24 Arecor Limited Protein formulation
US8476408B2 (en) 2008-06-13 2013-07-02 Alize Pharma Sas Unacylated ghrelin and analogs as therapeutic agents for vascular remodeling in diabetic patients and treatment of cardiovascular disease
EP2328607A1 (en) 2008-07-16 2011-06-08 Arecor Limited Stable formulation of a therapeutic protein
WO2011041584A2 (en) 2009-09-30 2011-04-07 President And Fellows Of Harvard College Methods for modulation of autophagy through the modulation of autophagy-enhancing gene products
KR20140097336A (en) 2011-11-23 2014-08-06 암젠 인크 Methods of treatment using an antibody against interferon gamma
LT2790721T (en) 2011-12-15 2019-02-11 Millendo Therapeutics Sas Fragments of unacylated ghrelin for use in the treatment of prader-willi syndrome
US9657062B2 (en) 2012-02-03 2017-05-23 Zealand Pharma A/S Ghrelin analogues
FR2987268B1 (en) * 2012-02-28 2014-07-11 Ammtek LIQUID FORMULATIONS OF HYPOGLYCEMIC SULFAMIDES
US20160122403A1 (en) 2013-07-16 2016-05-05 Liat Mintz Ghrelin splice variant for treating neuronal damage,neurodegenerative disease, parkinsons disease, alzheimers disease, and/or depression
US9119832B2 (en) 2014-02-05 2015-09-01 The Regents Of The University Of California Methods of treating mild brain injury
WO2016040224A1 (en) * 2014-09-09 2016-03-17 The Feinstein Institute For Medical Research Treatment of radiation injury using ghrelin
JP6580581B2 (en) * 2014-09-27 2019-09-25 大日本住友製薬株式会社 Pharmaceutical composition for injection
US20180296647A1 (en) * 2015-10-14 2018-10-18 Kurume University Prophylactic and therapeutic agent for rett syndrome (rtt) comprising ghrelin as active ingredient
US20170121385A1 (en) 2015-10-28 2017-05-04 Oxeia Biopharmaceuticals, Inc. Methods of treating neurodegenerative conditions
JP2018169273A (en) * 2017-03-29 2018-11-01 東ソー株式会社 Peptide adsorption inhibitor
JP7561325B2 (en) * 2018-07-03 2024-10-04 フェネック ファーマシューティカルズ，インコーポレイテッド Formulation of anhydrous sodium thiosulfate

Family Cites Families (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0651637B2 (en) * 1985-03-28 1994-07-06 エーザイ株式会社 Peptide adsorption prevention composition
JPS62238298A (en) * 1986-04-08 1987-10-19 Sanraku Inc New anthracycline antibiotics DCP-1 and 2
JPH0669956B2 (en) * 1988-09-30 1994-09-07 旭化成工業株式会社 Anti-adsorption agent for polypeptides
DE4126983A1 (en) * 1991-08-15 1993-02-18 Boehringer Mannheim Gmbh METHOD FOR THE PRODUCTION OF HUMAN-PROTEIN-CONTAINING, PRESERVED MEDICAMENTS FOR INFUSION OR INJECTION USE
SE9300105D0 (en) 1993-01-15 1993-01-15 Kabi Pharmacia Ab STABLE PROTEIN SOLUTION
US5482931A (en) * 1993-06-29 1996-01-09 Ferring Ab Stabilized pharmaceutical peptide compositions
DE4340781C3 (en) 1993-11-30 2000-01-27 Novartis Ag Liquid preparations containing cyclosporin and process for their preparation
TW426523B (en) 1995-04-06 2001-03-21 Hoffmann La Roche Interferon solution
US5843903A (en) * 1995-11-27 1998-12-01 The Administrators Of The Tulane Educational Fund Targeted cytotoxic anthracycline analogs
US5780431A (en) 1996-09-20 1998-07-14 Neurobiological Technologies, Inc. Pharmaceutical formulations of corticotropin releasing factor having improved stability in liquid form
US5998381A (en) * 1996-12-06 1999-12-07 Ophidian Pharmaceuticals, Inc. Compounds that bind bacterial pili
BR9913857A (en) 1998-09-17 2001-06-12 Lilly Co Eli Protein formulations
EP1795598B1 (en) * 1999-07-23 2009-11-18 Kenji Kangawa Novel peptides
US20020061838A1 (en) 2000-05-17 2002-05-23 Barton Holmquist Peptide pharmaceutical formulations
EP1286697A2 (en) * 2000-05-17 2003-03-05 Eli Lilly And Company Method for selectively inhibiting ghrelin action
JP2004513069A (en) 2000-05-19 2004-04-30 レストラゲン，インコーポレイテッド Peptide drug formulation
CA2411667A1 (en) 2000-05-30 2001-12-06 Merck & Co. Inc. Ghrelin analogs
DE60224284T2 (en) 2001-06-28 2008-12-18 Novo Nordisk A/S STABLE FORMULATION OF MODIFIED GLP-1
US7138489B2 (en) * 2002-04-11 2006-11-21 Daiichi Asubio Pharma Co., Ltd. Method for producing a modified peptide
CN100411683C (en) 2002-05-21 2008-08-20 阿斯比奥制药株式会社 Pharmaceutical composition containing ghrelin

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101541341B (en) * 2006-09-27 2013-06-05 益普生制药两合公司 Analogs of ghrelin substituted at the N-terminal
CN114306225A (en) * 2019-05-07 2022-04-12 前沿生物药业(南京)股份有限公司 Stable esbociclib composition

Also Published As

Publication number Publication date
JPWO2003097083A1 (en) 2005-10-06
US8518893B2 (en) 2013-08-27
WO2003097083A1 (en) 2003-11-27
US20120264691A1 (en) 2012-10-18
CA2471879C (en) 2014-02-25
IL162796A0 (en) 2005-11-20
KR20050012224A (en) 2005-01-31
JP5000848B2 (en) 2012-08-15
EP1506786A1 (en) 2005-02-16
AU2003235401A1 (en) 2003-12-02
EP1506786B1 (en) 2016-11-23
US20060166871A1 (en) 2006-07-27
US20080081787A1 (en) 2008-04-03
US8440628B2 (en) 2013-05-14
CN100411683C (en) 2008-08-20
ES2615484T3 (en) 2017-06-07
EP1506786A4 (en) 2009-07-29
IL162796A (en) 2011-01-31
BR0306685A (en) 2005-04-26
CA2471879A1 (en) 2003-11-27
AU2003235401B2 (en) 2008-09-25

Similar Documents

Publication Publication Date Title
CN1622822A (en) Medicinal compositions containing ghrelin
CN1051083C (en) Novel substituted purinyl derivatives with immunomodulating activity
CN1035256C (en) Preparation method of new polypeptide compound
CN1229390C (en) Peptide that lower blood glucose levels
CN1180839C (en) Powdery preparation for mucosal administration containing polymeric medicine
CN1635832A (en) Method for administering GLP-1 molecules
CN1832959A (en) Polyethelene glycol link glp-1 compounds
CN1596120A (en) Medicinal compositions for nasal absorption
CN1091314A (en) The pharmaceutical composition that contains calcitonin
CN1406124A (en) Nasally administrable cyclic peptide compositions
CN1703424A (en) GLP-1 derivatives and transmicosal absorption preparations thereof
CN1495198A (en) Analogs of glucagon-like peptide-1
CN1571796A (en) Melanocortin receptor ligands
CN1195776C (en) Treatmet of obesity
CN1176947C (en) Parathyroid hormone analogues for the treatment of osteoporosis
CN1925866A (en) Treatment of bacterial infections
CN1871020A (en) Antagonistic analogs of GH-RH (2003)
CN1861634A (en) Dog interferon alpha, preparation process and use thereof
CN1642570A (en) Stable pharmaceutical composition containing factor viii
CN1612748A (en) Remedies for dry eye and diseases associated with dry eye
CN1049166A (en) The replacement analogue of Magainin peptide
CN1732182A (en) Peptide and pharmaceutical composition containing said peptide
CN1612891A (en) Method for producing modified peptide
CN1121223C (en) Freeze-dried composition containing glycosphingolipid and process for producing the same
CN1781933A (en) Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C56 Change in the name or address of the patentee

Owner name: SANKYO CO.

Free format text: FORMER NAME: ASUBIO PHARMA CO., LTD.

CP03 Change of name, title or address

Address after: Tokyo, Japan

Co-patentee after: Kangawa, Kenji

Patentee after: DAIICHI SANKYO Co.,Ltd.

Address before: Tokyo, Japan

Co-patentee before: Kangawa, Kenji

Patentee before: ASUBIO PHARMA Co.,Ltd.

CX01 Expiry of patent term

Granted publication date: 20080820

CX01 Expiry of patent term